Determination of doxorubicin and doxorubicinol in plasma of cancer patients by high-performance liquid chromatography

Anal Biochem. 1999 Jan 15;266(2):216-21. doi: 10.1006/abio.1998.2943.


A high-performance liquid chromatographic assay with fluorescence detection has been developed for the determination of doxorubicin and its metabolite doxorubicinol in plasma of cancer patients. Quantitative extraction was achieved by a single protein-precipitation step of 1-ml samples with 500 microl of acetone in the presence of 100 microl of zinc sulfate [70% (w/v) in water]. Doxorubicin and doxorubicinol were separated isocratically on a column packed with Inertsil ODS-80A material and a mobile phase composed of water:acetonitrile:tetrahydrofuran (76:24:0.5, v/v/v). The related compound daunorubicin was used as internal standard. The column effluent was monitored fluorimetrically at an excitation wavelength of 480 nm and an emission wavelength of 560 nm, with a band width of 40 nm. The calibration graphs of doxorubicin and doxorubicinol were linear over a range of 1.0 to 100 and 0.50 to 50. 0 ng/mL, respectively, with lower limits of quantitation of 1.0 and 0.50 ng/ml. Results obtained from a 4-day validation study demonstrated excellent accuracy (91.0-106%) and precision (0.90-10. 2%) across the calibration ranges for both compounds. The developed method has been applied extensively to a clinical study to examine the pharmacokinetics and metabolism of doxorubicin in patients cotreated with a potent inhibitor of MDR1 P-glycoprotein activity, GF120918.

MeSH terms

  • Antineoplastic Agents / blood*
  • Antineoplastic Agents / pharmacokinetics
  • Chromatography, High Pressure Liquid / methods*
  • Doxorubicin / analogs & derivatives*
  • Doxorubicin / blood*
  • Doxorubicin / metabolism
  • Doxorubicin / pharmacokinetics
  • Female
  • Humans
  • Ovarian Neoplasms / blood*
  • Ovarian Neoplasms / metabolism
  • Sensitivity and Specificity
  • Time Factors


  • Antineoplastic Agents
  • Doxorubicin
  • adriamycinol